IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ mukherjee-2008-1 CASK Functions as a Mg2+-independent neurexin kinase. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu Cell (0092-8674) 2008 Only protein-protein interactions 4 imex curation mukherjee-2008-3 CASK Functions as a Mg2+-independent neurexin kinase. in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. Details of origin of Nrxn1 supplied by author konark.mukherjee@utsouthwestern.edu Cell (0092-8674) 2008 Only protein-protein interactions 4 imex curation mukherjee-2008-2 CASK Functions as a Mg2+-independent neurexin kinase. in vitro In vitro fluorescence spectr classical fluorescence spectroscopy fluorescence spectr predetermined predetermined participant predetermined Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. konark.mukherjee@utsouthwestern.edu Cell (0092-8674) 2008 Only protein-protein interactions 4 imex curation mukherjee-2008-4 CASK Functions as a Mg2+-independent neurexin kinase. in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting predetermined predetermined participant predetermined Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. The nucleotide was cross-linked with protein using UV-Stratalinker. konark.mukherjee@utsouthwestern.edu Cell (0092-8674) 2008 Only protein-protein interactions 4 imex curation mukherjee-2008-5 CASK Functions as a Mg2+-independent neurexin kinase. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. Details of origin of Nrxn1 supplied by author konark.mukherjee@utsouthwestern.edu Cell (0092-8674) 2008 Only protein-protein interactions 4 imex curation mukherjee-2008-6 CASK Functions as a Mg2+-independent neurexin kinase. in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Mukherjee K., Sharma M., Urlaub H., Bourenkov GP., Jahn R., Sudhof TC., Wahl MC. Synapse - Interactions of proteins with an established role in the presynapse. Details of origin of Nrxn1 supplied by author konark.mukherjee@utsouthwestern.edu Cell (0092-8674) 2008 Only protein-protein interactions 4 imex curation adenosine 3'-monophosphate adenosine 3'-monophosphate (3'-AMP) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) cskp_human Peripheral plasma membrane protein CASK CASK LIN2 Calcium/calmodulin-dependent serine protein kinase Protein lin-2 homolog protein protein human Homo sapiens Human MADDDVLFEDVYELCEVIGKGPFSVVRRCINRETGQQFAVKIVDVAKFTSSPGLSTEDLKREASICHMLKHPHIVELLETYSSDGMLYMVFEFMDGADLCFEIVKRADAGFVYSEAVASHYMRQILEALRYCHDNNIIHRDVKPHCVLLASKENSAPVKLGGFGVAIQLGESGLVAGGRVGTPHFMAPEVVKREPYGKPVDVWGCGVILFILLSGCLPFYGTKERLFEGIIKGKYKMNPRQWSHISESAKDLVRRMLMLDPAERITVYEALNHPWLKERDRYAYKIHLPETVEQLRKFNARRKLKGAVLAAVSSHKFNSFYGDPPEELPDFSEDPTSSGLLAAERAVSQVLDSLEEIHALTDCSEKDLDFLHSVFQDQHLHTLLDLYDKINTKSSPQIRNPPSDAVQRAKEVLEEISCYPENNDAKELKRILTQPHFMALLQTHDVVAHEVYSDEALRVTPPPTSPYLNGDSPESANGDMDMENVTRVRLVQFQKNTDEPMGITLKMNELNHCIVARIMHGGMIHRQGTLHVGDEIREINGISVANQTVEQLQKMLREMRGSITFKIVPSYRTQSSSCERDSPSTSRQSPANGHSSTNNSVSDLPSTTQPKGRQIYVRAQFEYDPAKDDLIPCKEAGIRFRVGDIIQIISKDDHNWWQGKLENSKNGTAGLIPSPELQEWRVACIAMEKTKQEQQASCTWFGKKKKQYKDKYLAKHNAVFDQLDLVTYEEVVKLPAFKRKTLVLLGAHGVGRRHIKNTLITKHPDRFAYPIPHTTRPPKKDEENGKNYYFVSHDQMMQDISNNEYLEYGSHEDAMYGTKLETIRKIHEQGLIAILDVEPQALKVLRTAEFAPFVVFIAAPTITPGLNEDESLQRLQKESDILQRTYAHYFDLTIINNEIDETIRHLEEAVELVCTAPQWVPVSWVY amp-pnp AMP-PNP small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) nrx1b_rat Neurexin-1-beta Nrxn1 Neurexin I-beta protein protein rat Rattus norvegicus (Rat) Rat MYQRMLRCGAELGSPGGGSSGGAGGRLALLWIVPLTLSGLLGVAWGASSLGAHHIHHFHGSSKHHSVPIAIYRSPASLRGGHAGTTYIFSKGGGQITYKWPPNDRPSTRADRLAIGFSTVQKEAVLVRVDSSSGLGDYLELHIHQGKIGVKFNVGTDDIAIEESNAIINDGKYHVVRFTRSGGNATLQVDSWPVIERYPAGNNDNERLAIARQRIPYRLGRVVDEWLLDKGRQLTIFNSQATIIIGGKEQGQPFQGQLSGLYYNGLKVLNMAAENDANIAIVGNVRLVGEVPSSMTTESTATAMQSEMSTSIMETTTTLATSTARRGKPPTKEPISQTTDDILVASAECPSDDEDIDPCEPSSGGLANPTRVGGREPYPGSAEVIRESSSTTGMVVGIVAAAALCILILLYAMYKYRNRDEGSYHVDESRNYISNSAQSNGAVVKEKQPSSAKSANKNKKNKDKEYYV tnp-atp 2',3'-O-(2,4,6- trinitrophenyl) adenosine 5'-triphosphate (TNP-ATP) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) ChEBI entry requested atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) cask-amp Crystallisation of CASK with 3'-AMP 739761 N/A 739767 unspecified role unspecified role neutral component neutral component N/A 739768 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false 1.85A working 20.2%, free 26.7% Fig. 1, Table S1, Supp. Fig. 3 cask-amppnp-1 Crystallisation of CASK with AMP-PNP 739761 N/A 739769 unspecified role unspecified role neutral component neutral component N/A 739768 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false 2.19A working 20.2%, free 25.7% Fig. 1B, 2B, Table S1, Supp. Fig. 2 cask-1 Autophosphorylation of CASK 739762 N/A 739768 putative self putative self putative self putative self region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false O14936:p.[Ser24Asp;Val26Leu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) phosphorylation phosphorylation reaction phosphorylation false true false KmATP of =0.563 mM, Vmax = 4.9 nmol/umol enzyme/min KiMg2+ = 61 uM Figs. 4A-D, Supp. Fig. S11A Phosphorylation of Ser-151 and Ser-155 identified by mass spectrometry Mg2+ and other divalent cations inhibited but did not abolish the autophosphorylation of CASK, presumably by reducing its ATP affinity cask-nrxn1-1 Phosphorylation of Nrxn1 by CASK 739762 N/A 739768 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false O14936:p.[Ser24Asp;Val26Leu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 739770 enzyme target enzyme target substrate substrate neutral component neutral component c-terminal tail sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) phosphorylation phosphorylation reaction phosphorylation false false false KmATP of neurexin-1 phosphorylation by CASK (=748.7 uM, Vmax = 159.4 nmol/umol enzyme/min Fig. 5, Supp. Figs. 10, 11B Phosphorylation of Nrxn1 identified by mass spectrometry cask-tnp-atp Interaction of CASK with TNP-ATP 739763 N/A 739768 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 739771 unspecified role unspecified role neutral component neutral component direct interaction direct interaction false false false Fig. 3A-D ATP titrations revealed that a 500-fold excess of ATP reduced the TNP-ATP fluorescence by 50% (KiATP = 0.365 mM), consistent with competitive binding of ATP and TNP-ATP to a single site that exhibits a much higher affinity for TNP-ATP than for ATP. Divalent cations could also inhibit this interaction, but the authors speculate that this was by complexing with ATP rather than by binding to the enzyme. EDTA stimulates the interaction, presumably by chelating divalent cations. cask-atp-1 Interacion of CASK with ATP 739764 N/A 739772 unspecified role unspecified role bait bait N/A 739768 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Supp. Fig. 6A ATP binding is inhibited by Mg2+ but not by Na+. cask-nrxn1-2 Colocalisation of Nrxn1 with CASK 739765 N/A 739768 unspecified role unspecified role neutral component neutral component region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 739770 unspecified role unspecified role neutral component neutral component c-terminal tail sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false cherry tag region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Supp. Fig. S8A coexpression of a CASK-EGFP fusion protein with neurexin-1β in HEK293 cells recruits CASK from the cytosol to the plasma membrane Q63373; O14936 transfection cask-nrxn1-3 Interaction of Nrxn1 with CASK 739766 N/A 739770 unspecified role unspecified role bait bait c-terminal tail sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) N/A 739768 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Supp. Fig. 9A Binding does not appear to be dependent on adenine-nucleotides